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Enhanced Delivery of Protein Fused to Cell Penetrating Peptides to Mammalian Cells
Jung-Il Moon1, Min-Joon Han2,#, Shin-Hye Yu3,#, Eun-Hye Lee4, Sang-Mi Kim4, Kyuboem Han3, Chang-Hwan Park4,5,6,*, Chun-Hyung Kim3
1Purdue Institute for Integrative Neuroscience, Purdue University,
2Department of Hematology, St. Jude Children’s Research Hospital,
3Paean Biotechnology Institute, Paean Biotechnology, Inc.,
4Hanyang Biomedical Research Institute and 5Department of Microbiology and 6Graduate School of Biomedical Science and Engineering, Hanyang University
Abstract
Recent progress in cellular reprogramming technology and lineage-specific cell differentiation has provided great opportunities for translational research. Because virus-based gene delivery is not a practical reprogramming protocol, protein-based reprogramming has been receiving attention as a safe way to generate reprogrammed cells. However, the poor efficiency of the cellular uptake of reprogramming proteins is still a major obstacle. Here, we reported key factors which improve the cellular uptake of these proteins. Purified red fluorescent proteins fused with 9xLysine (dsRED-9K) as a cell penetrating peptide were efficiently delivered into the diverse primary cells. Protein delivery was improved by the addition of amodiaquine. Furthermore, purified dsRED-9K was able to penetrate all cell lineages derived from mouse embryonic stem cells efficiently. Our data may provide important insights into the design of protein-based reprogramming or differentiation protocols.
Abstract, Accepted Manuscript(in press) [Submitted on August 28, 2018, Accepted on October 1, 2018]
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